末端補(bǔ)體復(fù)合體C5b-9(C5b-9)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)
ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9)
MAC; Membrane Attack Complex
- 編號(hào)SEC350Hu
- 物種Homo sapiens (Human,人) 相同的名稱(chēng),不同的物種。
- 實(shí)驗(yàn)方法雙抗夾心
- 反應(yīng)時(shí)長(zhǎng)3h
- 檢測(cè)范圍1.25-80ng/mL
- 靈敏度最小可檢測(cè)劑量小于等于0.52ng/mL.
- 樣本類(lèi)型serum, plasma and other biological fluids
- 下載 英文說(shuō)明書(shū) 中文說(shuō)明書(shū)
- 規(guī)格 48T96T 96T*5 96T*10 96T*100
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特異性
本試劑盒用于檢測(cè)末端補(bǔ)體復(fù)合體C5b-9(C5b-9),經(jīng)檢測(cè)與其它相似物質(zhì)無(wú)明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來(lái)源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。
回收率
分別于定值血清及血漿樣本中加入一定量的末端補(bǔ)體復(fù)合體C5b-9(C5b-9)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。
樣本 | 回收率范圍(%) | 平均回收率(%) |
serum(n=5) | 79-90 | 85 |
EDTA plasma(n=5) | 80-97 | 84 |
heparin plasma(n=5) | 92-105 | 101 |
精密度
精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
批內(nèi)差: CV<10%
批間差: CV<12%
線性
在定值血清及血漿樣本內(nèi)加入適量的末端補(bǔ)體復(fù)合體C5b-9(C5b-9),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中末端補(bǔ)體復(fù)合體C5b-9(C5b-9)含量的測(cè)定值與理論值的比率。
樣本 | 1:2 | 1:4 | 1:8 | 1:16 |
serum(n=5) | 80-91% | 88-95% | 81-90% | 78-102% |
EDTA plasma(n=5) | 81-99% | 91-102% | 94-105% | 98-105% |
heparin plasma(n=5) | 89-97% | 87-99% | 94-101% | 80-91% |
穩(wěn)定性
經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來(lái)進(jìn)行操作可減少人為誤差。
實(shí)驗(yàn)流程
1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
2. 加樣(標(biāo)準(zhǔn)品及樣本)100µL,37°C孵育1小時(shí);
3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
4. 洗板3次;
5. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
6. 洗板5次;
7. 加TMB底物90µL,37°C孵育10-20分鐘;
8. 加終止液50µL,立即450nm讀數(shù)。
實(shí)驗(yàn)原理
將末端補(bǔ)體復(fù)合體C5b-9(C5b-9)抗體包被于96孔微孔板中,制成固相載體,向微孔中分別加入標(biāo)準(zhǔn)品或標(biāo)本,其中的末端補(bǔ)體復(fù)合體C5b-9(C5b-9)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的末端補(bǔ)體復(fù)合體C5b-9(C5b-9)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標(biāo)記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過(guò)氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。顏色的深淺和樣品中的末端補(bǔ)體復(fù)合體C5b-9(C5b-9)呈正相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。
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相關(guān)產(chǎn)品
編號(hào) | 適用物種:Homo sapiens (Human,人) | 應(yīng)用(僅供研究使用,不用于臨床診斷!) |
SEC350Hu | 末端補(bǔ)體復(fù)合體C5b-9(C5b-9)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMC350Hu | 末端補(bǔ)體復(fù)合體C5b-9(C5b-9)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) | FLIA Kit for Antigen Detection. |
參考文獻(xiàn)
雜志 | 參考文獻(xiàn) |
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